LOCUS       sequence-105955-        7474 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses VfAU1-LOV domain (Vaucheria frigida Aureochrome 1) 
            preceded by mVenus, the extracellular and transmembrane domain of 
            p75NTR and a restriction site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7474)
  TITLE     MYR-VfAU1-LOV_135
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7474)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: mVenus-P75ECD-VfAU1-LOV_458
FEATURES             Location/Qualifiers
     source          1..7474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      943..952
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1039..1041,1042..1752)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1342..1361)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1689..1710
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2965..2991
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3068..3085)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3074..3298
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3344..3772
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3431..3450)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3641..3662
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3781..3801)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3786..4115
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3966..4101
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4028..4047
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4182..4976
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4236..4255)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4846..4865
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5150..5271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5187..5206)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5241..5260
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5320..5336)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5320..5336)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5333..5355)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5344..5360
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5368..5374,5375..5392,5393..5398))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5413..5434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5551..5568)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5722..6307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5802..5821)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6478..7269,7270..7338))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7101..7120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7339..7443)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgcggccggc gcgccaccat gggagcaggt
      961 gcaactggta gagcaatgga cggacctaga ctcctgctgc tcctcctgct cggagtgtcc
     1021 ctgggtggag caatcgatgt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1081 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1141 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     1201 gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac
     1261 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1321 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1381 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1441 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     1501 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1561 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1621 ctgcccgaca accactacct gagctaccag tccaaactga gcaaagaccc caacgagaag
     1681 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1741 gagctgtaca agatcgataa agaagcatgc ccaactggac tctacaccca ttctggagag
     1801 tgctgcaagg cctgcaatct gggtgaagga gtcgctcagc catgtggagc aaaccagacc
     1861 gtctgtgagc cttgcctgga ttctgtgaca ttctccgacg tggtgtctgc caccgagcct
     1921 tgcaagccat gcaccgagtg cgttggcctg cagtcaatgt ctgctccctg cgtggaggct
     1981 gacgacgcag tgtgtagatg cgcatatggc tactaccagg atgaaactac tgggaggtgc
     2041 gaggcatgta gggtgtgtga ggcaggcagt ggtctggttt tcagttgcca ggacaagcag
     2101 aacaccgtct gcgaggagtg tccagacggg acatacagcg acgaagccaa ccatgtcgat
     2161 ccctgcctgc catgtaccgt ttgcgaagat accgaacgcc agctgagaga atgcactaga
     2221 tgggctgacg ccgaatgtga ggagatccca ggacggtgga ttactcgctc tactcctcca
     2281 gaagggtcag atagcaccgc tcccagcacc caagaacccg aagctcctcc cgagcaggat
     2341 ctgattgcct ccacagttgc tggagtcgtg actaccgtga tgggttcctc acaacccgtc
     2401 gtgaccagag ggacaaccga caacctgatt ccagtgtatt gctctattct ggctgccgtc
     2461 gtggtgggac tggttgcata catagccttc aagagggcca ccggtggccc tgactacagt
     2521 ctcgtgaagg ctctgcaaat ggcacaacag aattttgtca ttacagacgc ctccctccca
     2581 gacaacccta tcgtctacgc cagtagaggg tttctgacac tgacaggcta ttctctcgac
     2641 cagatcctgg gcaggaactg caggtttctg caagggccag aaacagaccc aagagctgtg
     2701 gataagatca ggaatgccat caccaaaggc gttgatacca gtgtctgtct gctgaattat
     2761 agacaggatg gcacaacctt ctggaatctc ttcttcgtgg ctggactcag agattctaag
     2821 ggcaatattg tcaactacgt cggagtgcag tcaaaggtga gcgaagatta tgccaagctg
     2881 ctggtcaacg agcagaacat tgagtacaaa ggtgtgcgca ccagtaacat gctgcgcaga
     2941 aagcccggtg gatccggagt cgactatccg tacgacgtac cagactacgc actcgactaa
     3001 gaattccacc acactggact agtggatccg agctcggtac caagcttaag tttaaaccgc
     3061 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3121 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3181 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3241 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3301 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3361 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3421 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3481 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3541 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3601 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3661 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3721 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3781 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3841 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3901 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3961 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4021 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4081 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4141 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4201 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4261 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4321 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4381 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4441 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4501 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4561 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4621 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4681 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4741 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4801 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4861 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4921 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4981 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5041 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5101 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5161 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5221 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5281 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5341 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5401 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5461 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5521 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5581 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5641 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5701 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5761 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5821 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5881 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5941 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6001 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6061 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6121 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6181 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6241 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6301 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6361 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6421 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6481 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6541 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6601 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6661 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6721 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6781 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6841 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6901 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6961 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7021 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7081 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7141 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7201 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7261 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7321 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7381 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7441 gcgcacattt ccccgaaaag tgccacctga cgtc
//
